Cell Culture Contamination Part 2

Recent studies have shown that at least 20% of the animal cell cultures currently in use in the US are contaminated by either microorganisms or other cell lines! This three part live, on-line seminar series will review the scope of this major problem and examine some of its causes and techniques for avoiding it. It will also explore some key, easy to employ strategies for preventing these losses by careful culture management. This webinar will discuss good asceptic technique: developing a practical approach to asceptic technique; reducing day-to-day contamination problems in the lab; helpful hings for avoiding contamination.

From basal media with feeder layers or serum to highly defined recombinant growth factors, cytokine, and extracellular matrix, there are many ways to grow the same cell type. The choice is dependent on scale, cost, control, skill, and regulatory factors.

This webinar will cover:
- Different ways to grow the same cell type
- The actual material costs of various methods
- Methods used to optimize formulations

Speaker Bio:
Kevin Kelly graduated from Hawaii Pacific University and for 15 years worked on process scale-up and optimization for extracellular matrix proteins, growth factors, cytokines, antibodies, ELISA kits, and Corning® BioCoat™ products.

Currently he provides applications support for invasion, migration, permeability, transport, differentiation, and metabolism assays.

Corning was pleased to have recently sponsored a GEN webinar highlighting the latest techniques for 3D cell culture in cancer research.

The use of 3D cell cultures has been rising sharply in recent years from its initial introduction, over two decades ago. Because 3D cultures more accurately mimic the cellular environment, they can be used to study various forms of cancer by fostering the growth of organoids that replicate key properties of in vivo organ systems or the original tumors from which they were derived.

In addition, many 3D cultures are amenable to large-scale drug screens for rapid detection of phenotypic or genetic changes associated with therapeutic compounds—an approach that opens the door for the use of 3D culture as an integral part of personalized medicine.

In this GEN webinar, panelists discussed how the latest 3D cell culture methods have facilitated breakthroughs in their research projects.

Ömer H. Yilmaz, M.D., Ph.D., Assistant Professor of Biology at the Massachusetts Institute of Technology, will describe his work on how adult stem cells and their microenvironment adapt to diverse conditions within the context of tissue regeneration and cancer initiation through the use of ex vivo intestinal organoid assays.

Wojciech Senkowski, Doctoral candidate in the Department of Medical Sciences at Uppsala University in Sweden, will discuss his current work, which looks for applications of various tumor spheroid models in high-throughput drug screening for ways to identify novel compounds that target these cell populations

This presentation describes critical factors for selecting microcarriers, as well as starting protocols to help you optimize the attachment and expansion of cells in spinner flask and bioreactor environments.

In addition, you’ll learn about new dissolvable microcarrier technology, which provides unique advantages for cells that cannot be easily separated from standard microcarriers.

Speaker Bio:

Jennifer Weber is a senior development scientist with Corning Life Sciences. She has helped develop a variety of products for culturing advanced cell types including Corning® Synthemax™, a synthetic, xeno-free surface, and Corning stemgro ® hMSC, a serum-free, chemically defined medium for hMSC culture.

She recently transitioned to microcarrier product development and customer support for bioprocess applications. As part of this role, she facilitates customer adoption of Corning products for specific applications through protocol development, on-site technical support, and in-house customer-driven projects.

Cell culture media is required for successful and reproducible research but the catalog is full of acronyms and various formulation tables. Classical mammalian cell culture media formulations are very diverse both in terms of the number available and the concentration of constituents. Additionally, each medium was designed for specific cell types and culturing conditions.

Speaker Bio:
Brian Posey is a Product Development Manager for cell culture media at Corning Life Sciences. Brian has over 10 years experience in cell biology and industrial scale cGMP manufacturing of both liquid and powder cell culture media. Since joining Corning in 2012, Brian has led numerous innovative technology projects for the media business ranging from customer technology transfer for production scale-up to developing new serum-free media for industrial and stem cell lines.

Organic anion-transporting polypeptides (OATPs) play an important role in hepatic uptake of a variety of clinically important drugs. The significant differences in OATP/Oatp-dependent drug transport between human and preclinical species presents a challenge for interspecies extrapolation of drug pharmacodynamics and pharmacokinetics. The assessment of the difference in hepatic uptake between species using an in vitro model is highly desired to support mechanistic studies and to understand the differences observed between species in vivo.

The Corning® TransportoCells™ model has demonstrated significant value in terms of supporting in vitro assessment of drug interaction with SLC transporters in drug discovery and development. Recently, several animal species of Oatps were introduced into the Corning TransportoCells portfolio. This webinar will focus on the development of the newly available animal species and characterization of the differences in OATP/Oatp in substrate specificity and kinetics using this "thaw and go" model.

Three-dimensional (3D) cell culture models, which offer significant improvements over traditional two dimensional monolayer cell culture in terms of maintaining morphological and functional characteristics of tissue, are increasingly being incorporated in drug discovery as model systems to study disease or for screening for chemotherapeutic efficacy or drug toxicity. This presentation will demonstrate the use of Corning® spheroid microplates to culture and assay spheroids in a rapid and highly reproducible format that enables the formation of a single multi-cellular spheroid, centered in each well.

Corning spheroid microplates are multiple well, cell culture plates with opaque walls and unique clear, round well-bottom geometry that utilize the Corning Ultra-Low Attachment surface coating. The coating is hydrophilic, biologically inert and non-degradable. Representative data will be shown exhibiting the use of the spheroid microplate in more specialized assays including the formation of neurospheres from neural stem cells (NSCs), a valuable model to study neurogenesis and neural development, as well as analysis of NSC proliferation and migration. We will also demonstrate the formation of hepatospheres using Corning HepatoCells, an immortalized alternative to primary human hepatocytes, in combination with the SCREEN-WELL Hepatotoxicity library from Enzo Life Sciences for 3D hepatotoxicity screening. Finally, we will show the use of the Corning spheroid microplate to generate spheroids comprised of multiple cell types, demonstrating the impact that including multiple cell types in 3D assays can have on therapeutic outcome in a chemotherapeutic assay.

Although filtration is commonly practiced in many laboratories, it is a critical step in preparing media and buffers, clarifying samples, removing microbial contaminants, concentrating and inline filtering gases, to name a few applications. In order to choose the correct filter for your application, you need to consider the application, the material(s) to be filtered, and other factors that will be covered in this presentation. This webinar will focus on:

Dr. Mark Rothenberg graduated from Emory University with his Ph.D. in Cell and Developmental Biology. Over the past 25 years, Mark has held positions in both academia and industry where he has developed an expertise in the areas of assay development and cell culture. He currently holds the position of Manager Scientific Training and Education with Corning Life Sciences.

The importance of genetic variations in drug transporters for drug disposition and response has been increasingly recognized in the past decade. The drug transporter organic anion transporting polypeptide 1B1 (OATP1B1) is genetically polymorphic and plays a major role in hepatic uptake of a variety of clinically important drugs. Predicting the pharmacokinetic effect of these genetic variants on the drug disposition is critical for understanding the inter-individual variations to drug efficacy and safety. Corning TransportoCells was introduced to support in vitro assessment of drug interaction with SLC transporter. Specially, the newly developed TransportoCells OATP1B1 SNPs, alongside with OATP1B1 wild type, offer an in vitro tool kit for studying the potential impact of genetic variants on drug PK.

This webinar will focus on the development and characterization of the newly available TransportoCells OATP1B1 genetic variants, three animal species Oatp family members and other drug transporters of emerging importance in drug development.

Primary human hepatocytes (PHHs) and other hepatic cell models possess several limitations. For instance, PHH’s large lot-to-lot variation requires qualification tests with each lot, resulting in high costs and increased lead time. Furthermore, other non-primary hepatic cells can have insufficient fold induction in some lots and conditions.

This presentation will introduce Corning® HepatoCells for ADME/Tox studies. Derived from primary human hepatocytes, Corning HepatoCells are a renewable, hepatocyte-like cell line that retains most of the physiological properties of their parental hepatocytes such as mature hepatocyte-like morphology and induction response to prototypical inducers of CYP3A4, 1A2, and 2B6. Characterization of Corning HepatoCells for ADME/Tox studies will be presented, along with data demonstrating how the model system can be used for prediction of clinical CYP induction.

Achieving your ADME/Tox testing goals requires experience, quality data, and proper alignment with regulatory guidance. Failure to meet these important requirements can put your drug discovery and pre-clinical goals at risk.
This presentation will provide an informative overview of how you can advance and reach your pre-clinical drug discovery goals. It will discuss the importance of core contract research capabilities, including enzyme induction, enzyme inhibition, and transporter interaction. In addition, we will review new capabilities and opportunities including CYP induction and SLC transporter assay services – all designed to align with regulatory agency guidance documents.

Speaker Bio:
David Stresser is the Program Manager of Corning® Gentest℠ Contract Research Services at Corning Life Sciences since 2001, having held prior positions of Product Manager and Study Director since joining Corning in 1998. Prior to this, he was a post-doctoral associate in the laboratory of David Kupfer at the University of Massachusetts Medical School in Worcester, Massachusetts. He did his graduate work in the laboratory of David E. Williams at Oregon State University in Corvallis, Oregon receiving a Ph.D. in toxicology in 1994. Dr. Stresser has authored or co-authored 40 articles or book chapters in the field of drug metabolism and has been an invited speaker at various national and international meetings, pharmaceutical companies, and universities.

Join us on June 22nd for a special Corning-sponsored webinar presented by ATCC®.

Abstract:
Animal cell lines are important in vitro systems and tools for scientists in diverse disciplines such as basic cell biology, genetic mapping, gene expression and gene therapy. Cell line authentication and characterization are crucial in these activities, yet they are underappreciated by most research scientists. Over the years numerous cell lines have been shown to be misidentified due, in part, to poor techniques and inadequate verification of cell line authenticity. Technological advances have given rise to improved capabilities. Cell line authentication now requires a comprehensive strategy that employs several complementary technologies for systematic testing for morphology, microbial contaminations, cellular identity/cross-contamination as well as functionality. The validity of conclusions drawn from research data is dependent on consistent and unequivocal verification of cell line identity and function. It is estimated that the financial loss incurred by poorly characterized or misidentified cell lines is in the millions of dollars. An overview of the current technologies used to authenticate cell lines will be presented.

Speaker Bio:
Dr. Yvonne A. Reid joined ATCC in 1980 and during the mid-1980’s her research focused on the use of DNA hypervariable probes for the intraspecies identification of cell lines. The evolution of this work has led to the implementation of routine screening of all human cell lines by STR analysis. She co-chaired the ATCC SDO committee on the Development Consensus Standard on the Authentication of Human Cell Lines: Standardization of STR profiling. Dr. Reid has more than 30 years of experience in cell biology, immunology and molecular biology. As Collection Scientist for the Cell Biology Program for over 10 years, she was responsible for acquisition of new animal cell lines and hybridomas into the Cell Biology General Collection.

This webinar will introduce the history, theory, basic techniques, and potential pit-falls of mammalian cell culture. It is designed for students and new lab technicians, as well as bench scientists interested in updating their techniques or knowledge in the field.

Topics to be discussed include:
• History and practical theories of cell culture and its impact on today’s science
• The requirements needed to set up a cell culture laboratory
• Challenges when performing mammalian cell culture and how to overcome them

About our Presenter:
Dr. Mark Rothenberg graduated from Emory University with his Ph.D. in Cell and Developmental Biology. Over the past 25 years, Mark has held positions in both academia and industry where he has developed an expertise in the areas of assay development and cell culture. He currently holds the position of Manager Scientific Training and Education with Corning Life Sciences.

Abstract:
There is a great interest in application of human mesenchymal stem cells (hMSCs) in cell therapy and tissue engineering due to their self-renewal, multi-lineage differentiation, immunomodulation, and trophic potential. One of the challenges faced in the clinical application of hMSCs is the need for efficient expansion of these cells in vitro without altering their capacity. Serum-free mammalian cell culture media, in particular, require optimization of the expansion protocols. Even subtle changes in routine handling can have a significant impact on the cells’ potential.

This seminar will cover the variables that can influence the desired regenerative and differentiation properties including medium selection, vessel surface treatment, impact of the cell source, and seeding density. We will also discuss how users can select the correct conditions for optimized growth and functionality.

Speaker Biography:
Brian Posey is a Product Development Manager for cell culture media at Corning Life Sciences. Brian has over 10 years experience in cell biology and industrial scale cGMP manufacturing of both liquid and powder cell culture media. Since joining Corning in 2012, Brian has lead numerous innovative technology projects for the media business ranging from customer technology transfer for production scale-up to developing new serum-free media for industrial and stem cell lines.

Abstract:
As a key determinant of drug pharmacokinetics, transporter mediated drug-drug interaction has garnered significant attention from the pharmaceutical industry and regulatory authorities. Corning offers a comprehensive list of tools to support drug transporter studies and recently introduced Corning® TransportoCells™ products to support in vitro assessment of drug interaction with SLC transporters. This new model provides a convenient “thaw and go” cell-based model with robust activity and consistent performance. In this webinar, we will provide an overview of Corning TransportoCells products along with applications for in vitro-to-in vivo correlation. Validation data will also be presented for the newly available TransportoCells products, including OATP1A2, OATP2B1, PEPT1, PEPT2, and NTCP.

Presenter Biography:
Dr. Na Li received her B.S degree in Biology from Fudan University, Shanghai, China, and her Ph.D. in Pharmacology from Dartmouth Medical School, Hanover, NH. Her major research focus is on drug transporters, including interspecies differences in hepatobiliary transporters, transporter quantification, and in vitro-to-in vivo extrapolation of drug pharmacokinetics. At Corning, Dr. Li contributes her expertise in in vitro drug transporter technology and its application in drug ADME.

Join us on January 28th for a special Corning-sponsored webinar presented by ATCC®.

Abstract:
The significance of 3D tissue modeling opens up new possibilities for the study of complex physiological processes in vitro. Advances in cell isolation, media development, substrates, and growth surfaces are leading to culture environments that provide better biological and functional properties than traditional 2D cell culture. These models may provide a more predictive analysis and result in a more streamlined process of drug discovery and development. In this webinar, we will discuss recent developments in 3D modeling using ATCC primary and hTERT immortalized cells with specialized Corning® permeable support culture systems in dermatologic and respiratory studies.

Presenter Biography:
Dr. Yukari Tokuyama is a Field Application Scientist at ATCC. Prior to this role, she led the Stem Cell Product Development group and focused on products for human induced pluripotent stem cells and lineage specific differentiation. She earned her Ph.D. in Cell and Molecular Biology from the College of Medicine at the University of Cincinnati, where she studied the mechanism of genomic instability in cancer. She completed her post-doctoral training at the Oregon Health & Science University, Oregon National Primate Research Center, with a research focus on human and non-human primate stem cell biology.

Join us for a special Corning sponsored webinar presented by Promega Corporation.

Cells cultured in 3D model systems often acquire relatively large in vivo-like structures compared to the thickness of a 2D monolayer of cells grown on standard plastic plates. Multicellular 3D culture systems containing more than one cell type and exhibiting formation of a complex extracellular matrix represent a more physiologically relevant environment, yet provide a challenge for assay chemistries originally designed for measuring events from monolayers of cells. There is an unmet need for guidelines for design and verification of convenient and effective assays useful for larger 3D microtissues. Critical factors to consider for each model system and cell type include effective penetration of detection reagents and/or complete lysis of microtissue structures using combinations of detergent and physical disruption. We will present the approach used to verify performance of a bioluminescent ATP detection assay for measuring cell viability, a caspase assay for detecting apoptosis, and cell stress reporter assays to detect mechanisms leading to cytotoxicity. Recommendations for factors to consider when verifying performance of cell health assays on 3D culture models will be presented.

Speaker Bio:

Dr. Terry Riss started the Cell Biology program at Promega Corporation in 1990 and has since held several R&D and Project Management positions. Dr. Riss managed development of cell viability, cytotoxicity, apoptosis, and protease assay systems and also led efforts to identify and promote multiplexing of cell-based assays to determine the mechanism of cell death. Dr. Riss now serves as Senior Product Specialist, Cell Health involved in outreach educational training activities including validating assay systems applied to 3D cell culture models.

Angiogenesis is the process by which a new blood supply is established from pre-existing blood vessels. It is initiated by degradation of vessel basement membrane, endothelial cell proliferation, invasion, and directional migration towards chemoattractants, tube formation, and finally the establishment of a new vasculature.

We will discuss a portfolio of Corning products designed to investigate specific stages of angiogenesis through standardized and quantitative in vitro cell-based assays.

Paula Flaherty is a Technology Manager at Corning Life Sciences. Her team develops strategy and products focused on the modulation of in vitro cell behavior using extracellular matrix, media, vessel design, and growth factors. Prior to joining Corning Life Sciences, Paula studied retinal degeneration at the Berman-Gund Laboratory, Harvard Medical School in Boston, MA. She received her bachelor’s degree in Microbiology from the State University of New York and is an In Vitro Cell Biology Fellow, W. Alton Jones Cell Science Center in Lake Placid, NY.

Are you relying on biochemical and cell based in vitro assays in your research? In this webinar, Dr. Mark Rothenberg will share “insider” tips and tricks for enhancing your biochemical and cell-based assay performance.
Critical parameters for achieving success with these assays include understanding the assay dynamics, the instrumentation involved in reading the assay, and, in the case of cell-based assays, the environment in which the cells are grown impacts cellular physiology. Factors such as microplate geometry, density, surfaces, and instrumentation all play important roles in the success of the assay.
Learn how to:
•Select the correct microplate and its role in the success of the assay
•Choose the correct tip for your liquid handling needs
•Determine the correct instrumentation and settings to run your assay

In the past several decades, the usage of mammalian cell culture processes have allowed for better understanding of basic research and manufacturing of higher quality products compared to previous methods. Although much progress has been developed throughout the years, limitations and challenges during optimization of such processes in scale-up of mammalian cell cultures may be encountered.

This webinar will provide a simple guide and review of novel cell culture vessels and surface technologies that have enabled researchers to improve suspension and adherent mammalian cell culture scale-up. Additionally, this webinar will cover certain parameters to consider when scaling-up cells. Selecting the correct vessel and cell culture conditions will increase throughput without increasing laboratory space, incubator space, or time.

The Corning Scientific Seminar Series is a series of free, online technical presentations that provide novel tips, best practices and proven techniques to help advance your research. Delivered by scientists to scientists, these one-hour broadcasts offer useful information and tips for lab technicians and researchers.